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Combining a Simple Method for DNA/RNA/Protein Co-Purification and Arabidopsis Protoplast Assay to Facilitate Viroid Research.

Viruses. 2019; 
Jiang J, Ma J, Liu B, Wang Y.
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Monoclonal Antibody Services After 10 min incubation with Rapidblock solution (VWR), the following specific primary antibodies were applied for overnight incubation at 4 ◦C: anti-GFP (Genscript, Piscataway, NJ) 1:2000, anti-Histone H3 (Genscript) 1:2000 dilution, anti-beta tubulin (Genscript) 1:2000 dilution, the polyclonal antibodies (MilliporeSigma) against Mitogen-activated protein kinase 3 (MAPK3) 1:1000, the monoclonal antibody (8WG16; Thermo Fisher Scientific) against the largest subunit of RNA polymerase II (NRPB1) 1:500, and the polyclonal antibodies (Aviva Systems Biology, San Diego, CA) against ribosomal protein L5 (RPL5) 1:1000. Get A Quote

摘要

Plant-viroid interactions represent a valuable model for delineating structure-function relationships of noncoding RNAs. For various functional studies, it is desirable to minimize sample variations by using DNA, RNA, and proteins co-purified from the same samples. Currently, most of the co-purification protocols rely on TRI Reagent (Trizol as a common representative) and require protein precipitation and dissolving steps, which render difficulties in experimental handling and high-throughput analyses. Here, we established a simple and robust method to minimize the precipitation steps and yield ready-to-use RNA and protein in solutions. This method can be applied to samples in small quantities, such as protopla... More

关键词

DNA/RNA/protein co-purification; protoplast; viroid